Analyzing Lung Inflammation and Tissue Changes in Mice

We performed bronchoalveolar lavage (BAL) in mice to analyze the lung inflammatory response and sacrificed them on days 14 and 21 to analyze the lung tissues. For BAL, the mice were anesthetized, a catheter was intubated into the upper respiratory tract, and 0.9 mL of PBS was administered twice to obtain the BAL fluid (BALF). Cells obtained from BALF were stained with Diff-Quik (Systmex, Kobe, Japan), and the numbers of macrophages, neutrophils, eosinophils, and lymphocytes within the sample were counted. Lung tissue samples were fixed and embedded in paraffin and stained with H&E, MT, terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL), and periodic acid-Schiff (PAS) stains. Inflammation, fibrosis, apoptosis, and mucus production in the lungs were semi-quantified and scored according to previous studies [43 (link)–46 (link)].

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Choi S.M., Mo Y., Bang J.Y., Ko Y.G., Ahn Y.H., Kim H.Y., Koh J., Yim J.J, & Kang H.R. (2023). Classical monocyte-derived macrophages as therapeutic targets of umbilical cord mesenchymal stem cells: comparison of intratracheal and intravenous administration in a mouse model of pulmonary fibrosis. Respiratory Research, 24, 68.

Publication 2023

Apoptosis Bronchoalveolar lavage Bronchoalveolar lavage fluid Catheter Cells Deoxynucleotidyl transferase Dutp Embedded paraffin Eosinophils Fibrosis Inflammation Lung inflammatory Lungs Lymphocytes Macrophages Mice Mucus Neutrophils Periodic acid Respiratory tract Stains Tissue

Corresponding Organization :

Other organizations : Seoul National University Hospital, Seoul National University

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Variable analysis

independent variables

Day of sacrifice (14 and 21)

dependent variables

Number of macrophages, neutrophils, eosinophils, and lymphocytes in bronchoalveolar lavage fluid (BALF)
Inflammation in lung tissues
Fibrosis in lung tissues
Apoptosis in lung tissues
Mucus production in lung tissues

control variables

Anesthesia of mice
Intubation of catheter into upper respiratory tract
Administration of 0.9 mL PBS twice to obtain BALF
Staining of cells from BALF with Diff-Quik
Fixation and paraffin embedding of lung tissue samples
Staining of lung tissue samples with H&E, MT, TUNEL, and PAS

controls

No explicit mention of positive or negative controls

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